Rapid and selective cloning of monitor peptide, a novel cholecystokinin-releasing peptide, using minimal amino acid sequence and the polymerase chain reaction.
cDNA transcripts encoding rat monitor peptide (MP) have been cloned from a lambda-ZAP-II phage library using minimal specific amino acid sequence (six residues), the polymerase chain reaction (PCR), and multivalent PCR probes to distinguish MP transcripts from those that encode a closely related peptide, pancreatic secretory trypsin inhibitor. DNA sequence analysis of 3 cDNA transcripts, MP1-3, revealed the complete amino acid sequence of the prepeptide (79 residues) including an 18-residue hydrophobic signal sequence at the NH2 terminus. Sequence divergence in both coding and 3' noncoding regions indicates a potential exon-exon junction with alternative splicing, which results in a truncated peptide with Arg 58 at the COOH terminus as well as alternative selection of poly(A) signals, respectively. The 5' nontranslated region of MP1 mRNA (282 nucleotides (nt] contains four upstream ATGs. Conserved structure between MP and anionic trypsinogen mRNAs within 9 nt immediately upstream of the AUG initiation codon may be involved in coupling the expression of MP with anionic trypsinogen, a condition which appears to be required to monitor the intake of dietary protein in the rat.
Fukuoka
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Laboratory of Cell and Molecular Biology, Charles A. Dana Research Institute, Beth Israel Hospital, Boston, Massachusetts.
Scheele
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GA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002766:Cholecystokinin / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015723:Gene Library; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010179:Pancreas / Q000502:physiology; D016133:Polymerase Chain Reaction; D051381:Rats; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000235:genetics*; D014361:Trypsin Inhibitors / Q000235:genetics*
D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D015345:Oligonucleotide Probes; C504705:Spink1 protein, rat; D014361:Trypsin Inhibitors; D014359:Trypsin Inhibitor, Kazal Pancreatic; D004247:DNA; D002766:Cholecystokinin